Canine CD34: Cloning of the cDNA and evaluation of an antiserum to recombinant protein

Peter A. McSweeney, Katherine A. Rouleau, Rainer Storb, Laura Bolles, Philip M. Wallace, Mary Beauchamp, Ljiljana Krizanac-Bengez, Peter F Moore, George Sale, Brenda Sandmaier, Thierry De Revel, Frederick R. Appelbaum, Richard A. Nash

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Increasingly, enriched populations of hematopoietic progenitors are used in experimental and clinical transplantation studies. The separation of progenitors is based on the expression of CD34, a marker preferentially expressed on progenitor cells. The dog model has been important for preclinical transplant studies, because it has proven predictive for outcomes in human hematopoietic stem cell transplantation. To identify and isolate canine hematopoietic progenitors, we have cloned a cDNA encoding a CD34 homologue from a canine myelomonocytic leukemia cell line, ML2. The CD34 homologue cDNA predicts an amino acid sequence that is highly conserved with human and murine CD34 in the cytoplasmic domain, transmembrane domain, and C- terminal end of the extracellular domain, but shows considerable divergence from these sequences at the amino-terminal end of the protein. In Western blotting studies, canine CD34 homologue (caCD34) appears to be a heavily and variably glycosylated protein with a molecular weight of approximately 100 kD and shows some tissue-specific differences in protein mass. To evaluate the expression of caCD34 protein, the extracellular domain of caCD34 was expressed as an Ig fusion protein and used as an immunogen to generate a rabbit polyclonal antiserum. The antiserum reacted against the fusion protein, against vascular endothelium, and with three leukemic cell lines. Approximately 1% of canine bone marrow cells stained brightly with antibodies to caCD34 and this population was 25- to 50-fold enriched for colony-forming units-granulocyte-macrophage as compared to unfractionated marrow mononuclear cells. These findings suggest that the canine CD34 homologue is expressed on bone marrow progenitor cells and, thus, that this molecule should be a valuable marker for identifying and isolating canine hematopoietic progenitors for experimental hematopoiesis and stem cell transplantation.

Original languageEnglish (US)
Pages (from-to)1992-2003
Number of pages12
JournalBlood
Volume88
Issue number6
StatePublished - Sep 15 1996

Fingerprint

Cloning
Recombinant Proteins
Canidae
Organism Cloning
Immune Sera
Complementary DNA
Proteins
Cells
Stem cells
Bone
Fusion reactions
Bone Marrow Cells
Transplants
Macrophages
Stem Cells
Cell Line
Granulocyte-Macrophage Progenitor Cells
Hematopoietic Stem Cell Transplantation
Molecular weight
Hematopoiesis

ASJC Scopus subject areas

  • Hematology

Cite this

McSweeney, P. A., Rouleau, K. A., Storb, R., Bolles, L., Wallace, P. M., Beauchamp, M., ... Nash, R. A. (1996). Canine CD34: Cloning of the cDNA and evaluation of an antiserum to recombinant protein. Blood, 88(6), 1992-2003.

Canine CD34 : Cloning of the cDNA and evaluation of an antiserum to recombinant protein. / McSweeney, Peter A.; Rouleau, Katherine A.; Storb, Rainer; Bolles, Laura; Wallace, Philip M.; Beauchamp, Mary; Krizanac-Bengez, Ljiljana; Moore, Peter F; Sale, George; Sandmaier, Brenda; De Revel, Thierry; Appelbaum, Frederick R.; Nash, Richard A.

In: Blood, Vol. 88, No. 6, 15.09.1996, p. 1992-2003.

Research output: Contribution to journalArticle

McSweeney, PA, Rouleau, KA, Storb, R, Bolles, L, Wallace, PM, Beauchamp, M, Krizanac-Bengez, L, Moore, PF, Sale, G, Sandmaier, B, De Revel, T, Appelbaum, FR & Nash, RA 1996, 'Canine CD34: Cloning of the cDNA and evaluation of an antiserum to recombinant protein', Blood, vol. 88, no. 6, pp. 1992-2003.
McSweeney PA, Rouleau KA, Storb R, Bolles L, Wallace PM, Beauchamp M et al. Canine CD34: Cloning of the cDNA and evaluation of an antiserum to recombinant protein. Blood. 1996 Sep 15;88(6):1992-2003.
McSweeney, Peter A. ; Rouleau, Katherine A. ; Storb, Rainer ; Bolles, Laura ; Wallace, Philip M. ; Beauchamp, Mary ; Krizanac-Bengez, Ljiljana ; Moore, Peter F ; Sale, George ; Sandmaier, Brenda ; De Revel, Thierry ; Appelbaum, Frederick R. ; Nash, Richard A. / Canine CD34 : Cloning of the cDNA and evaluation of an antiserum to recombinant protein. In: Blood. 1996 ; Vol. 88, No. 6. pp. 1992-2003.
@article{5cc8b05aaece4316822469eba0e00e94,
title = "Canine CD34: Cloning of the cDNA and evaluation of an antiserum to recombinant protein",
abstract = "Increasingly, enriched populations of hematopoietic progenitors are used in experimental and clinical transplantation studies. The separation of progenitors is based on the expression of CD34, a marker preferentially expressed on progenitor cells. The dog model has been important for preclinical transplant studies, because it has proven predictive for outcomes in human hematopoietic stem cell transplantation. To identify and isolate canine hematopoietic progenitors, we have cloned a cDNA encoding a CD34 homologue from a canine myelomonocytic leukemia cell line, ML2. The CD34 homologue cDNA predicts an amino acid sequence that is highly conserved with human and murine CD34 in the cytoplasmic domain, transmembrane domain, and C- terminal end of the extracellular domain, but shows considerable divergence from these sequences at the amino-terminal end of the protein. In Western blotting studies, canine CD34 homologue (caCD34) appears to be a heavily and variably glycosylated protein with a molecular weight of approximately 100 kD and shows some tissue-specific differences in protein mass. To evaluate the expression of caCD34 protein, the extracellular domain of caCD34 was expressed as an Ig fusion protein and used as an immunogen to generate a rabbit polyclonal antiserum. The antiserum reacted against the fusion protein, against vascular endothelium, and with three leukemic cell lines. Approximately 1{\%} of canine bone marrow cells stained brightly with antibodies to caCD34 and this population was 25- to 50-fold enriched for colony-forming units-granulocyte-macrophage as compared to unfractionated marrow mononuclear cells. These findings suggest that the canine CD34 homologue is expressed on bone marrow progenitor cells and, thus, that this molecule should be a valuable marker for identifying and isolating canine hematopoietic progenitors for experimental hematopoiesis and stem cell transplantation.",
author = "McSweeney, {Peter A.} and Rouleau, {Katherine A.} and Rainer Storb and Laura Bolles and Wallace, {Philip M.} and Mary Beauchamp and Ljiljana Krizanac-Bengez and Moore, {Peter F} and George Sale and Brenda Sandmaier and {De Revel}, Thierry and Appelbaum, {Frederick R.} and Nash, {Richard A.}",
year = "1996",
month = "9",
day = "15",
language = "English (US)",
volume = "88",
pages = "1992--2003",
journal = "Blood",
issn = "0006-4971",
publisher = "American Society of Hematology",
number = "6",

}

TY - JOUR

T1 - Canine CD34

T2 - Cloning of the cDNA and evaluation of an antiserum to recombinant protein

AU - McSweeney, Peter A.

AU - Rouleau, Katherine A.

AU - Storb, Rainer

AU - Bolles, Laura

AU - Wallace, Philip M.

AU - Beauchamp, Mary

AU - Krizanac-Bengez, Ljiljana

AU - Moore, Peter F

AU - Sale, George

AU - Sandmaier, Brenda

AU - De Revel, Thierry

AU - Appelbaum, Frederick R.

AU - Nash, Richard A.

PY - 1996/9/15

Y1 - 1996/9/15

N2 - Increasingly, enriched populations of hematopoietic progenitors are used in experimental and clinical transplantation studies. The separation of progenitors is based on the expression of CD34, a marker preferentially expressed on progenitor cells. The dog model has been important for preclinical transplant studies, because it has proven predictive for outcomes in human hematopoietic stem cell transplantation. To identify and isolate canine hematopoietic progenitors, we have cloned a cDNA encoding a CD34 homologue from a canine myelomonocytic leukemia cell line, ML2. The CD34 homologue cDNA predicts an amino acid sequence that is highly conserved with human and murine CD34 in the cytoplasmic domain, transmembrane domain, and C- terminal end of the extracellular domain, but shows considerable divergence from these sequences at the amino-terminal end of the protein. In Western blotting studies, canine CD34 homologue (caCD34) appears to be a heavily and variably glycosylated protein with a molecular weight of approximately 100 kD and shows some tissue-specific differences in protein mass. To evaluate the expression of caCD34 protein, the extracellular domain of caCD34 was expressed as an Ig fusion protein and used as an immunogen to generate a rabbit polyclonal antiserum. The antiserum reacted against the fusion protein, against vascular endothelium, and with three leukemic cell lines. Approximately 1% of canine bone marrow cells stained brightly with antibodies to caCD34 and this population was 25- to 50-fold enriched for colony-forming units-granulocyte-macrophage as compared to unfractionated marrow mononuclear cells. These findings suggest that the canine CD34 homologue is expressed on bone marrow progenitor cells and, thus, that this molecule should be a valuable marker for identifying and isolating canine hematopoietic progenitors for experimental hematopoiesis and stem cell transplantation.

AB - Increasingly, enriched populations of hematopoietic progenitors are used in experimental and clinical transplantation studies. The separation of progenitors is based on the expression of CD34, a marker preferentially expressed on progenitor cells. The dog model has been important for preclinical transplant studies, because it has proven predictive for outcomes in human hematopoietic stem cell transplantation. To identify and isolate canine hematopoietic progenitors, we have cloned a cDNA encoding a CD34 homologue from a canine myelomonocytic leukemia cell line, ML2. The CD34 homologue cDNA predicts an amino acid sequence that is highly conserved with human and murine CD34 in the cytoplasmic domain, transmembrane domain, and C- terminal end of the extracellular domain, but shows considerable divergence from these sequences at the amino-terminal end of the protein. In Western blotting studies, canine CD34 homologue (caCD34) appears to be a heavily and variably glycosylated protein with a molecular weight of approximately 100 kD and shows some tissue-specific differences in protein mass. To evaluate the expression of caCD34 protein, the extracellular domain of caCD34 was expressed as an Ig fusion protein and used as an immunogen to generate a rabbit polyclonal antiserum. The antiserum reacted against the fusion protein, against vascular endothelium, and with three leukemic cell lines. Approximately 1% of canine bone marrow cells stained brightly with antibodies to caCD34 and this population was 25- to 50-fold enriched for colony-forming units-granulocyte-macrophage as compared to unfractionated marrow mononuclear cells. These findings suggest that the canine CD34 homologue is expressed on bone marrow progenitor cells and, thus, that this molecule should be a valuable marker for identifying and isolating canine hematopoietic progenitors for experimental hematopoiesis and stem cell transplantation.

UR - http://www.scopus.com/inward/record.url?scp=9544237125&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=9544237125&partnerID=8YFLogxK

M3 - Article

C2 - 8822918

AN - SCOPUS:9544237125

VL - 88

SP - 1992

EP - 2003

JO - Blood

JF - Blood

SN - 0006-4971

IS - 6

ER -